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1.
Gomperts BD 《The Biochemical journal》1967,102(3):782-790
1. Whole blood was incubated at 37°, while being dialysed against a large volume of iso-osmotic bicarbonate buffer, pH7·4. The buffer contained glucose and the essential inorganic components of blood plasma in proportion. 2. After 3hr. of incubation in vitro there is a loss of red-cell 2,3-diphosphoglycerate. 3. Isotope experiments show that this is due to an accelerated rate of destruction of this compound. 4. Simultaneously, there is an increase in the median of red-cell osmotic fragility. 5. After extended periods of incubation there is a decrease in the metabolic rate and a decrease in the ratio of the rates of lactate production to glucose consumption. 6. There is a continuous loss of total adenine nucleotide and, after the first 12hr. of incubation, a tendency for the intracellular Na+ and K+ to equilibrate with the plasma. 7. The standard deviation of red-cell osmotic fragility expressed among the red-cell population increases exponentially with the time of incubation. 相似文献
2.
Mast cells permeabilized by streptolysin O secrete histamine and lysosomal enzymes in response to provision of a dual effector system comprising Ca2+ and a guanine nucleotide (e.g., GTP-gamma-S2) at concentrations in the micromolar range. These are both necessary and together they are sufficient. There is no requirement for adenosine triphosphate (ATP) and hence no obligatory phosphorylation reaction in the terminal stages of the exocytotic pathway. When exocytosis is induced by Ca2(+)-plus-GTP-gamma-S (i.e., no ATP) added at times after permeabilization (the permeabilization interval), cellular responsiveness declines so that there is no response to provision of the two effectors (both at 10(-5)M) if they are initially withheld and then added after 5 min. Here we show that this decline in responsiveness is characterized by a time-dependent reduction in the effective affinity for Ca2+. Affinity for Ca2+ and hence secretory competence can then be restored if ATP is added alongside the stimulus. Unlike cells stimulated to secrete at the time of permeabilization, exocytosis from cells that have undergone the cycle of permeabilization-induced refractoriness followed by ATP-induced restoration can be triggered by Ca2+ alone: after such conditioning there is no requirement for guanine nucleotide. In contrast, dependence on guanine nucleotide remains mandatory in cells that have been pretreated (i.e., before permeabilization) with okadaic acid (understood to be an inhibitor of protein phosphatases 1 and 2A) or phorbol myristate acetate (an activator of protein kinase C). These results indicate that obligatory dependence on guanine nucleotide is retained when the cells are treated under conditions conducive to maintained phosphorylation. It is concluded that the exocytotic mechanism of permeabilized mast cells is enabled by a dephosphorylation reaction and that the effector of the guanosine triphosphate (GTP)-binding protein (G epsilon) that mediates exocytosis is likely to be a protein phosphate. 相似文献
3.
Genetic variation at the Major Histocompatibility Complex locus DQ beta was
analyzed in 233 beluga whales (Delphinapterus leucas) from seven
populations: St. Lawrence Estuary, eastern Beaufort Sea, eastern Chukchi
Sea, western Hudson Bay, eastern Hudson Bay, southeastern Baffin Island,
and High Arctic and in 12 narwhals (Monodon monoceros) sympatric with the
High Arctic beluga population. Variation was assessed by amplification of
the exon coding for the peptide binding region via the polymerase chain
reaction, followed by either cloning and DNA sequencing or single-stranded
conformation polymorphism analysis. Five alleles were found across the
beluga populations and one in the narwhal. Pairwise comparisons of these
alleles showed a 5:1 ratio of nonsynonymous to synonymous substitutions per
site leading to eight amino acid differences, five of which were
nonconservative substitutions, centered around positions previously shown
to be important for peptide binding. Although the amount of allelic
variation is low when compared with terrestrial mammals, the nature of the
substitutions in the peptide binding sites indicates an important role for
the DQ beta locus in the cellular immune response of beluga whales.
Comparisons of allele frequencies among populations show the High Arctic
population to be different (P < or = .005) from the other beluga
populations surveyed. In these other populations an allele, Dele-DQ
beta*0101-2, was found in 98% of the animals, while in the High Arctic it
was found in only 52% of the animals. Two other alleles were found at high
frequencies in the High Arctic population, one being very similar to the
single allele found in narwhal.
相似文献
4.
Plasma-membrane location of phosphatidylinositol hydrolysis in rabbit neutrophils stimulated with formylmethionyl-leucylphenylalanine. 总被引:4,自引:3,他引:1 下载免费PDF全文
Rabbit peritoneal neutrophils, disrupted by sonication, were separated into three subcellular fractions by sucrose-step-gradient centrifugation and these were analysed with respect to biochemical markers. They comprised a high-speed supernatant containing the cytosol, a light particulate fraction enriched in Golgi and plasma membranes and a heavy particulate fraction enriched in granules and nuclei. The light particulate fraction was further separated into its components, which were identified as Golgi membranes (galactosyltransferase activity) and plasma membranes ((radioactivity derived from labelling intact cells with [125I]di-iodosulphanilic acid diazonium salt and [3H]formylmethionyl-leucylphenylalanine ([3H]fMet-Leu-Phe) binding)). In cells prelabelled with [3H]glycerol, the hydrolysis of phosphatidylinositol due to cell stimulation with fMet-Leu-Phe (10 nM) was shown to occur in the light particulate fraction. The [32P]Pi-labelling of phosphatidate, which is an early consequence of phosphatidylinositol hydrolysis, also occurred in this fraction. Analytical sucrose-gradient centrifugation of the light particulate fraction showed that the stimulated increment in [32P]phosphatidate (and thus by implication the initial phosphatidylinositol breakdown) was localized in the plasma membrane. 相似文献
5.
The role of cadherins during primordial germ cell migration and early gonad formation in the mouse 总被引:6,自引:0,他引:6
Bendel-Stenzel MR Gomperts M Anderson R Heasman J Wylie C 《Mechanisms of development》2000,91(1-2):143-152
Primordial germ cells (PGCs) are the founder cells of the gametes. In mammals, PGCs migrate from the hindgut to the genital ridges, where they coalesce with each other and with somatic cells to form the primary sex cords. We show here that, in both sexes, PGCs express P- and E-cadherins during and after migration, and N-cadherin at post-migratory stages. E-Cadherin is not expressed by PGCs whilst in the hindgut, but is upregulated as they leave. Blocking antibodies against E-, but not P-cadherin cause defective PGC-PGC coalescence, and in some cases, ectopic PGCs. 相似文献
6.
Elongation factor-1 alpha occurs as two copies in bees: implications for phylogenetic analysis of EF-1 alpha sequences in insects 总被引:5,自引:1,他引:4
We report the complete sequence of a paralogous copy of elongation factor-1
alpha (EF-1 alpha) in the honeybee, Apis mellifera (Hymenoptera: Apidae).
This copy differs from a previously described copy in the positions of five
introns and in 25% of the nucleotide sites in the coding regions. The
existence of two paralogous copies of EF-1 alpha in Drosophila and Apis
suggests that two copies of EF-1 alpha may be widespread in the
holometabolous insect orders. To distinguish between a single, ancient gene
duplication and parallel, independent fly and bee gene duplications, we
performed a phylogenetic analysis of hexapod EF-1 alpha sequences.
Unweighted parsimony analysis of nucleotide sequences suggests an ancient
gene duplication event, whereas weighted parsimony analysis of nucleotides
and unweighted parsimony analysis of amino acids suggests the contrary:
that EF-1 alpha underwent parallel gene duplications in the Diptera and the
Hymenoptera. The hypothesis of parallel gene duplication is supported both
by congruence among nucleotide and amino acid data sets and by
topology-dependent permutation tail probability (T-PTP) tests. The
resulting tree topologies are also congruent with current views on the
relationships among the holometabolous orders included in this study
(Diptera, Hymenoptera, and Lepidoptera). More sequences, from diverse
orders of holometabolous insects, will be needed to more accurately assess
the historical patterns of gene duplication in EF-1 alpha.
相似文献
7.
Ahmed E. Hegab Vi Luan Ha Yasser S. Attiga Derek W. Nickerson Brigitte N. Gomperts 《Journal of visualized experiments : JoVE》2012,(67)
The large airways are directly in contact with the environment and therefore susceptible to injury from toxins and infectious agents that we breath in 1. The large airways therefore require an efficient repair mechanism to protect our bodies. This repair process occurs from stem cells in the airways and isolating these stem cells from the airways is important for understanding the mechanisms of repair and regeneration. It is also important for understanding abnormal repair that can lead to airway diseases 2. The goal of this method is to isolate a novel stem cell population from the mouse tracheal submucosal gland ducts and to place these cells in in vitro and in vivo model systems to identify the mechanisms of repair and regeneration of the submucosal glands 3. This production shows methods that can be used to isolate and assay the duct and basal stem cells from the large airways 3.This will allow us to study diseases of the airway, such as cystic fibrosis, asthma and chronic obstructive pulmonary disease. Currently, there are no methods for isolation of submucosal gland duct cells and there are no in vivo models to study the regeneration of submucosal glands. 相似文献
8.
Bastien Gomperts Frédéric Lantelme Reinhard Stock 《The Journal of membrane biology》1970,3(1):241-266
Summary (1) When salts are added to buffered suspensions of membrane fragments containing the fluorochrome 1-anilino-8-naphthalenesulfonate (ANS), there is an increased fluorescence. This is caused by increased binding of the fluorochrome; the intrinsic fluorescence characteristics of the bound dye remain unaltered. These properties make ANS a sensitive and versatile indicator of ion association equilibria with membranes. (2) Alkali metal and alkylammonium cations bind to membranes in a unique manner. Cs+ binds most strongly to rat brain microsomal material, with the other alkali metals in the order Cs+>Rb+>K+>Na+>Li+. The reaction is endothermic and entropy driven. Monovalent cations are displaced by other monovalent cations. Divalent cations and some drugs (e. g., cocaine) displace monovalent cations more strongly. (3) Divalent cations bind to membranes (and to lecithin micelles) at four distinct sites, having apparent association constants between 50 and 0.2mm
–1. The characteristics of the titration suggest that only one species of binding site is present at any one time, and open the possibility that structural transitions of the unassociated coordination sites may be induced by divalent cation binding. Divalent cation binding at the weakest site (like monovalent cation binding) is endothermic and entropy driven. At the next stronger site, the reaction is exothermic. Monovalent cations affect divalent cation binding by reducing the activity coefficient: they do not appear to displace divalent cations from their binding sites. 相似文献
9.
This study continues analysis from a companion paper on over 350,000 insured Swedish dogs up to 10 years of age contributing
to more than one million dog-years at risk during 1995–2000. The age patterns for total and diagnostic mortality and for general
causes of death (trauma, tumour, locomotor, heart and neurological) are presented for numerous breeds. Survival estimates
at five, eight and 10 years of age are calculated. Survival to 10 years of age was 75% or more in Labrador and golden retrievers,
miniature and toy poodles and miniature dachshunds and lowest in Irish wolfhounds (91% dead by 10 years). Multivariable analysis
was used to estimate the relative risk for general and more specific causes of death between breeds accounting for gender
and age effects, including two-way interactions. Older females had tumour as a designated cause of death more often than males
in most breeds, but not in the Bernese mountain dog. Information presented in this and the companion paper inform our understanding
of the population level burden of disease, and support decision-making at the population and individual level about health
promotion efforts and treatment and prognosis of disease events. 相似文献
10.
Fabian Kloosterman Thomas J. Davidson Stephen N. Gomperts Stuart P. Layton Gregory Hale David P. Nguyen Matthew A. Wilson 《Journal of visualized experiments : JoVE》2009,(26)
Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. Lightweight recording devices carrying a high density array of tetrodes allow for the simultaneous monitoring of the activity of tens to hundreds of individual neurons. Here we describe a protocol for the fabrication of a micro-drive array with twenty one independently movable micro-drives. This device has been used successfully to record from hippocampal and cortical neurons in our lab. We show how to prepare a custom designed, 3-D printed plastic base that will hold the micro-drives. We demonstrate how to construct the individual micro-drives and how to assemble the complete micro-drive array. Further preparation of the drive array for surgical implantation, such as the fabrication of tetrodes, loading of tetrodes into the drive array and gold-plating, is covered in a subsequent video article.Open in a separate windowClick here to view.(88M, flv) 相似文献